Method for deactivating a contaminant

ABSTRACT

Methods, compositions and a kit for deactivating spills or leaks of HIV infected blood or anticancer drugs by applying to the leak or spill an aqueous solution containing calcium hypochlorite or sodium hypochlorite as the active ingredient. In order to thicken the aqueous solution and thus keep it from spreading beyond its intended area of application, the solution of calcium hypochlorite contains methylcellulose. In addition to chemically deactivating the active anticancer drug, the solution of the invention also effectively decolorizes it, thus preventing permanent stains on any surface or fabric with which the anticancer drug comes into contact.

CROSS-REFERENCE TO RELATED APPLICATION

This application is a divisional, pursuant to 37 C.F.R. §1.53(b), ofparent application Ser. No. 07/973,211, filed on Nov. 6, 1992 andentitled "METHOD AND COMPOSITION FOR DEACTIVATING HIV INFECTED BLOOD ANDFOR DEACTIVATING AND DECOLORIZING ANTICANCER DRUGS", now U.S. Pat. No.5,811,113, which is a continuation of application Ser. No 07/788,157,filed on Nov. 6, 1991 (now abandoned), which is a continuation-in-partof application Ser. No. 07/715,722, filed on Jun. 14, 1991 (nowabandoned), which is a continuation of application Ser. No. 07/377,062,filed Jul. 10, 1989, now abandoned, which is a continuation-in-part ofapplication Ser. No. 07/344,213, filed on Apr. 27, 1989 now abandoned,and the benefit of such earlier filing dates is hereby asserted pursuantto 35 U.S.C. §120.

BACKGROUND OF THE INVENTION

This invention relates to methods and compositions for deactivating HIV(human immunodeficiency virus) infected blood and anticancer drugs whichmay have been accidentally spilled or which have leaked from a patient,thus constituting a possible hazard to attending personnel. In addition,some anticancer drugs are inherently colored and such leakages or spillsmay stain clothes. The invention is effective to eliminate such stainsby decolorizing the drug.

Among the known antineoplastic or anticancer drugs which are used fortreating cancers of various types, some are known or suspected to be inthemselves carcinogenic. In addition, some of these anticancer drugs arealso dyes capable of creating unsightly stains on clothes and otherfabrics such as bedsheets. Accordingly, when solutions of anticancerdrugs are spilled or leaked, a possible hazard to attending personnel,as well as an unsightly stain on clothing and the like, may be created.

BRIEF DESCRIPTION OF THE INVENTION

In accordance with the invention, spills or leaks of HIV infected bloodand/or anticancer drugs are chemically deactivated by applying to theleak or spill an aqueous solution containing sodium or calciumhypochlorite as the active ingredient. In one embodiment, in order tothicken the aqueous solution and thus keep it from spreading beyond itsintended area of application, the hypochlorite solution containsmethylcellulose. In addition to chemically deactivating the activeanticancer drug, the composition of the invention also effectivelydecolorizes it, thus preventing permanent stains on any surface orfabric with which the anticancer drug comes into contact.

In a second embodiment of the present invention, spills or leaks of HIVinfected blood and/or anticancer drugs on stainless steel or ceramicwork surfaces vulnerable to etching by the corrosive alkaline sodium orcalcium hypochlorite solutions are inactivated with a two-step, towletteswabbing kit and process. A first absorbent, fibrous (preferably cottonand/or synthetic blend) towlette, impregnated with calcium hypochloriteor, preferably, sodium hypochlorite, is swabbed over the spilled HIVinfected blood- or drug-containing work surface to deactivate thespilled compound. A second absorbent, fibrous (preferably cotton and/orsynthetic blend) towlette, impregnated with sodium thiosulfate, is thenswabbed over the spilled HIV infected blood- or drug-containing worksurface to neutralize the alkaline calcium or sodium hypochloriteresidue from the first towlette, so as to prevent the alkaline residuefrom corroding or etching the work surface.

DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

In accordance with the invention, HIV infected blood, or any of a numberof active anticancer drugs, can be inactivated by applying thereto anaqueous solution containing 4-40% by weight of calcium or sodiumhypochlorite. HIV infected blood, as well as any anticancer agent whichcan undergo oxidative or alkaline degradation can be deactivated inaccordance with the method of the invention. Among the anticancer agentswhich can be effectively deactivated are the following:

anthracyclines, such as doxorubicin, daunorubicin, and epirubicin

anthracenes, such as mitoxantrone and bisantrene

alkylating agents, such as mitomycin-c, melphalan, cyclophosphamide,ifosphamide, thio-TEPA, decarbazine, carmustine, cisplatin, andcarboplatin

antimetabolites, such as fluorouracil, cytarabine, methotrexate, andmercaptopurine

biologicals, such as α-interferon, interleukin 2, tumor necrosis factor,G-CSF, and GM-CSF

miscellaneous compounds, such as vincristine, vinblastine, actinomycinD, bleomycin, etoposide, L-asparaginase, mAMSA, vindesine, andteniposide

In addition to chemically deactivating the anti-cancer drugs, the methodof the invention is also effective to decolorize those, e.g., theanthracyclines, doxorubicin and daunorubicin, which are naturally red incolor, and the alkylating agent mitomycin C, which is naturally purplein color, which stain fabrics which they contact. The method of theinvention renders the naturally colored anticancer drugs colorless, thuspreventing permanent damage to clothes or other materials which come incontact with the drugs. It has been found that deactivation andcolorization of the anticancer drug occur within approximately 30-60seconds after a suitable solution of hypochlorite is applied inaccordance with the invention.

The hypochlorite solution can be applied in any convenient manner,suitably by means of a manually operated spray bottle or othermechanical spray device. It is also within the contemplation of theinvention to use, in addition to such mechanical devices, an aerosoldispenser activated by a conventional aerosol propellant.

If the HIV infected blood and/or the drugs are spilled on a ceramic orstainless steel work surface, which surfaces may be vulnerable topermanent etching or disfiguration by the alkaline deactivator compoundscalcium hypochlorite and sodium hypochlorite, it is preferred to use atwo step, towlette swabbing kit and procedure to deactivate the spilledmaterial. A first absorbent, fibrous (preferably cotton and/or syntheticblend) towlette is impregnated with an aqueous solution containing4-40%, but preferably less than 6%, of a deactivator compound such ascalcium hypochlorite or, preferably, sodium hypochlorite by applying tothe first towlette an excess of aqueous deactivator solution, beyondthat amount necessary to completely saturate the first towlette. Theratio of deactivator volume to towlette weight should be in the range ofabout 1.8 to 15 ml deactivator per gram towlette (preferably 6.5 to 15ml/gm); alternatively, the ratio of deactivator volume to towlettesurface area should be in the range of about 0.01 to 0.2 ml deactivatorper cm² of towlette. The deactivator-impregnated towlette is wiped orswabbed over the spilled HIV infected blood- or drug-containing worksurface to deactivate the spilled compound; if the spilled compound is adrug which creates a stain, deactivation is indicated by disappearanceof the stain after swabbing.

A second absorbent, fibrous (preferably cotton and/or synthetic blend)towlette, impregnated with an aqueous solution of 4-40%, but preferablyless than 6%, of sodium thiosulfate, by applying to the second towlettean excess of the aqueous sodium thiosulfate solution, beyond that amountnecessary to completely saturate the second towlette. The ratio ofsodium thiosulfate volume to towlette weight should be in the range ofabout 1.8 to 15 ml deactivator per gram towlette (preferably 6.5 to 15ml/gm); alternatively, the ratio of sodium thiosulfate volume totowlette surface area should be in the range of about 0.01 to 0.2 mldeactivator per cm² of towlette. The sodium thiosulfate-impregnatedtowlette is then wiped or swabbed over the spilled HIV infected blood-or drug-containing work surface to neutralize the alkaline calcium orsodium hypochlorite residue from the first towlette, so as to preventthe alkaline residue from corroding or etching the work surface.Furthermore, sodium thiosulfate itself is capable of deactivatingseveral carcinogenic anticancer drugs, such as the alkylating agentsnitrogen mustard and cisplatin.

Suitable towlettes include "Terri® Wipers", nylon-reinforced 4 ply,#34770, each measuring 17×23 cm and weighing approximately 2.6 gram,manufactured by Kimberly Clark, Neenah, Wis., in addition to the thefollowing "Texwipe®" towlettes, manufactured by Texwipe Corporation,Upper Saddle River, N.J.: Technicon A, Absorbanal Applicators, and TX801Applicators.

The compatability of the various "Texwipe®" towlettes with sodiumhypochlorite was tested using the following procedure. Strips of thedifferent Texwipe® fabrics were cut into 1×1 inch squares and weighed.The fabric strips were then placed in 15 ml plastic vials. 5 ml of anaqueous solution containing 5.25% by weight of sodium hypochlorite wasadded to each vial. The vials were capped and incubated at roomtemperature for 10 days. The strips were then removed from the vials,air dried, and weighed. To test the deactivation ability of the sodiumhypochlorite solution remaining in each vial, 0.1 ml of a solutioncontaining 1.0 mg/ml of the drug doxorubicin was applied to each ofseveral "Terri® Wipers", nylon-reinforced 4 ply, #34770, each measuring17×23 cm and weighing approximately 2.6 gram, manufactured by KimberlyClark, Neenah, Wis. 0.1 ml samples of the sodium hypochlorite solutionremaining in each vial were then added to a respectivedoxorubicin-containing Terri® Wiper. Deactivation was inferred by acolor change from red to colorless.

The results of the "Texwipe®" tests are summarized in table A. Of thefour "Texwipe®" towlettes tested, Technicon A yielded the best results,although the bleach was completely inactivated by the Technicon fabricafter seven weeks. Two of the towlettes (Absorbanal Applicators andTX801 Applicators) exhibited significant weight gain after storage inthe bleach, indicating possible covalent bonding of the hypochloritedirectly to the fabric. The results for those three towlettes suggestthat it may be preferable to add excess solution to the towlettes toassure adequate deactivating activity after storage. A fourth "Texwipe®"towlette (TX811) was completely dissolved by the bleach, and is thus notsuitable for this application. Preferably, the sodium hypochlorite andsodium thiosulfate impregnated towlettes are prepared in advance andstored in separate sealed foil packages or the like until use.

                                      TABLE A                                     __________________________________________________________________________    TEXWIPE SAMPLE TESTS: Compatibility with Sodium Hypochlorite                                                  Activity with                                              Weights            respect to                                                                           Dry   % Difference                     Sample A  No.                                                                              (g) Quality  Appearance                                                                          Doxorubicin                                                                          Weight                                                                              In Weight                        __________________________________________________________________________    Technicon 1) .0674                                                                             Paper-like texture;                                                                    Clear Inactive                                                                             .0619  +8                              A         2) .0462                                                                             Tears easily                                                                           solution     .0472  +2                                        3) .0501                     .0460  -9/2                                                                   Average                                                                              +0.6                            Absorbanal Applicator                                                                   1) .420                                                                              Thick, soft; Tears                                                                     Yellow                                                                              Highly active                                                                        .1025 +144                             B         2) .511                                                                              with difficulty                                                                        solution     .1238 +142                                       3) .544                      .1234 +123                                                                    Average                                                                             +118                             TX811 Applicator                                                                        1) .0266                                                                             Paper-like quality;                                                                    Clear Slowly active;                                                                       None                                   D         2) .0966                                                                             very thin                                                                              solution                                                                            Incomplete                                                                           Available                                                              inactivation;                                           3) .0286              No wipes left                                 TX801 Applicator                                                                        1) .1128                                                                             Soft-life but sturdy                                                                   Yellow                                                                              Highly active                                                                        .1901  +88                             D         2) .0966        solution     .1582  +64                                       3) .1031                     .1733  +68                                                                    Average                                                                              +67                             __________________________________________________________________________     Bleach: National Sanitary Supply 5.25% w/v Sodium Hypochlorite                *All white cloth, single thickness; Approximate sample size 1 × 1       inch.                                                                    

It has also been determined that HIV infected blood and most anticancerdrugs can be deactivated by application of a solution of calcium orsodium hypochlorite in water at a concentration of 4% by weight. A fewof the anticancer drugs, notably mitoxantrone, may require the use of asolution having a substantially higher concentration of hypochlorite, upto 40% by weight, in order to achieve substantially completedeactivation of the anticancer drug.

In order to maintain the hypochlorite solution in contact with the drug,and to avoid displacement, it is desirable to increase the viscosity ofthe solution of hypochlorite. It has been found, however, thatconventional thickeners tend to deactivate the solution of hypochloriteat different rates. Thus, gelatin causes an immediate violentendothermic reaction when added to a solution of calcium hypochlorite,while polyvinylpyrolidine and polyethylene glycol cause deactivation ofthe hypochlorite solution within one hour. Methyl cellulose deactivatesthe hypochlorite, but at relatively slow rates so thathypochlorite-methylcellulose solutions maintain a practical degree ofeffectiveness for a reasonable time (e.g., 8 hours) after mixing.

In addition to deactivating of the hypochlorite, addition of a thickenerto a solution of hypochlorite causes an interaction between thehypochlorite and the thickener which reduces the ability of thethickener to increase the viscosity of the solution. Thus, for example,the viscosity of a typical solution of calcium hypochlorite andmethylcellulose drops by more than two-thirds after a period of threedays.

While the deactivating ability of the hypochlorite and the thickeningability of the thickeners decrease most rapidly in aqueous solution,similar effects occur when mixtures of the dry powders are stored. Ithas been found that mixtures of calcium hypochlorite and methylcellulosepowders stored in a dry condition lose a significant proportion ofdeactivating ability within 5 days and became almost completely inactiveafter 30 days at room temperatures, such that complete inactivation canoccur after 10 days at 50° C.

From the above, it is apparent that mixtures of hypochlorite andthickener, whether in solution or in dry form, have very limited shelflives. The solutions of the invention should be prepared freshimmediately before use and should not be stored for any extended periodsof time. Accordingly, the invention is suitably prepared and utilized asa two-package system including a first package containing an appropriatequantity of the hypochlorite, and a second package containing anappropriate quantity of the hypochlorite, and a second packagecontaining an appropriate quantity of the thickener, the contents of thepackage to be dissolved in a predetermined volume of water immediatelybefore use. In order to maintain the appropriate concentrations ofhypochlorite and thickener in the finished solution, i.e., 4-40% byweight of hypochlorite and 1-5% by weight of thickener, the proportionby weight of hypochlorite in the first package to the weight ofthickener in the second package is in the range of 1:1 to 40:1, andpreferably 1.2:1 to 2.0:1.

Methylcellulose maintains its ability to thicken the deactivatingsolutions of the invention for relatively long periods of time with arelatively small effect on the deactivating ability of the hypochlorite.Accordingly, methylcellulose is the preferred thickener for use in theinvention. Other common thickeners, such as agar, polyvinylpyrolidine,polyethylene glycol, and gelatin, either react violently when mixed withthe hypochlorite or produce solutions having too short an effective life(minutes to hours) for practical use.

The effectiveness of the method and composition of the invention aredemonstrated in the following examples.

EXAMPLE 1 Mutagenic Activity

Solutions of DNA intercalators (doxorubicin, daunorubicin andmitoxantrone) in concentrations of 10 ng/ml D5W were applied in 5 mlquantities to separate petri dishes. Two different concentrations ofcalcium hypochlorite, 40 g/l and 423 g/l, were applied in 1 ml. volumesseparately to the anticancer drug solutions in each petri dish for aperiod of one minute. After thorough mixing and neutralization of the pHto 7.4 using HCl, 1 ml of each petri dish solution was applied to 35 mmpetri dishes containing cultures of Salmonella Typhimurium TA 98 strain.The petri dishes containing bacteria plus overlaid mixtures of calciumhypochlorite/anticancer drug and control plates with bacteria alone werethen incubated at 37° C. for 24 hours. The colonies of revertantorganisms were counted and reported in Table 1.

                  TABLE 1                                                         ______________________________________                                        Mutagenic Activity from DNA Intercalators Treated                             with Calcium Hypochlorite                                                                No. Colonies of Revertant Organisms                                DNA Intercalator                                                                           Calcium Hypochlorite Concentration                               (10 ng/ml    None        40 g/l 423 g/l                                       ______________________________________                                        Doxorubicin  257         0      0                                             Daunorubicin 365         0      0                                             Mitoxantrone 164         44     0                                             None (control)                                                                              20         0      0                                             ______________________________________                                    

Table 1 reports the number of colonies of revertant (mutated) bacterialorganisms resulting from calcium hypochlorite treatment at two differentconcentrations of three different anticancer drugs. The controlbacterial plates (i.e., untreated with anticancer drug or calciumhypochlorite) contained only 20 bacterial colonies after 24 hours. Onthe other hand, the bacterial plates treated with the anticancer drug inthe absence of calcium hypochlorite contained between 164 and 365bacterial colonies per plate. At the higher concentration of calciumhypochlorite there were no colonies of revertant organisms in thebacterial plates and at the lower concentration of calcium hypochloriteonly mitoxantrone was associated with bacterial revertant growth.

EXAMPLE 2 Anthracene or Anthracycline Chromophore Recovery by HighPerformance Liquid Chromatography

Treatment of the drugs with calcium hypochlorite was carried out asdescribed in Example 1. One ml aliquots of doxorubicin, daunorubicin andmitoxantrone, separately, were taken from the petri dishes after calciumhypochlorite exposure at two different concentrations for a period ofone minute. Ten (10) ng of each of the anticancer drug (based on thefinal concentration resulting from the addition of calcium hypochloriteto the separate petri dishes containing each anticancer agent) was addedto a high performance liquid chromatography column and standard HPLCassays were carried out. A reverse phase C-18 bonded phase HPLCprocedure was used. The mobile phase consisted of 30:70 CH₃ CN:ammoniumacetate at a flow rate of 2 ml/min., using a Varian model 5020 HPLCunit. The anticancer drugs were detected using excitation at 480 nm andemission at 550 nm with a Schoeffel model FS 970 fluoroescence detector.At the higher concentration of calcium hypochlorite (423 g/l) thetreatment resulted in complete chemical degradation of all threeanticancer drugs. At the lower concentration, there was no chemicalevidence of either doxorubicin or daunorubicin and approximately 60%degradation of mitoxantrone.

                  TABLE II                                                        ______________________________________                                        Anthracene or Anthracycline Chromophore Recovery                              by High Performance Liquid Chromatography                                                Concentration ng by Fluroescence*                                  DNA Intercalator                                                                           Calcium Hypochlorite Concentration                               (10 ng on Column)                                                                          0-(Control)  40 g/l 423 g/l                                      ______________________________________                                        Doxorubicin  9.7          0      0                                            Daunomycin   9.8          0      0                                            Mitoxantrone 10.1         3.8    0                                            ______________________________________                                         *Reverse phase C18 bonded phase, mobile phase of 30:70 CH.sub.3 CN:           ammonium acetate, flow rate 2 ml/min, (Varian Model 5020); detection usin     excitation at 480 nm and emission at 550 nm (Schoeffel Model FS 970)     

EXAMPLE 3 Compatibility of Hypochlorite and Methylcellulose in AqueousSolution

The compatibility of calcium hypochlorite with methylcellulose inaqueous solution was investigated in a series of tests. Freshly madesolutions containing 4% by weight of calcium hypochlorite and 2.5% byweight of methylcellulose were examined for viscosity, pH, and abilityto deactivate anticancer drugs. Measurements of pH were made by acommercial pH meter while viscosity was measured by an instrument usingthe falling ball method. The ability to inactivate anticancer drugs wasevaluated by titration of stock solutions of doxorubicin (2 mg/ml) andmitoxantrone (2 mg/ml). The tests were performed by titrating theinactivating solution onto absorbent paper containing measured amountsof each anticancer agent. The end point was the number of drops ofinactivating solution required for complete neutralization of the redcolor of doxorubicin or the blue color of mitoxantrone.

The viscosity, pH, and neutralizing ability of the solutions weremeasured at intervals over a period of three to four days. The freshlymade mixture had an initial viscosity of 32 cps, a pH of 11.0, and anassigned value of 100% inactivating activity. These values changedrapidly after the solution was prepared. The viscosity dropped by almost40% in the first 12 hours, and by 68% within 72 hours, the pH valuedropped to 7.0 in one day, and the inactivating activity dropped to 50%within a period of 12-48 hours, depending on how thoroughly the solidingredients were dissolved in the mixture. Those compositions in whichthe components were not thoroughly dissolved retained inactivatingactivity for longer periods of time. By contrast, solutions in which thehypochlorite and methylcellulose were thoroughly dissolved became moreinactive at a faster rate. In general, a loss of 10-20% of activityoccurred within 8 hours after the aqueous solution was prepared.Accordingly, provided the solution was used within 8 hours ofpreparation, most of the inactivating ability of the solution wasretained.

EXAMPLE 4 Stability of Dry Mixtures of Calcium Hypochlorite andMethylcellulose

The stability of mixtures of calcium hypochlorite and methylcellulose indry form was tested by preparing individual mixtures of 4 grams ofcalcium hypochlorite and 2.5 grams of methylcellulose, and storing themixtures in plastic bottles at room temperature (25° C.) and at elevatedtemperature (50° C.). At various times, water was added to the mixtureof dry ingredients in an amount sufficient to produce 100 ml. of asolution containing 4 grams of calcium hypochlorite and 2.5 grams ofmethylcellulose. The solution was examined for pH, viscosity, andinactivating activity as described above. The results are given in TableIII.

                  TABLE III                                                       ______________________________________                                        Stability of Calcium Hypochlorite                                             and Methylcellulose Powders                                                               Properties of Solution**                                          Days of  Tempera-          Viscosity                                                                            Inactivating                                Storage  ture °C.)                                                                       pH       (cps)  Activity*                                   ______________________________________                                         1       25       11       30     100                                          5       25       10.5     28     85                                           7       25       10       27     71                                          10       25       8.5      18     60                                          14       25       7.0      5.7    45                                          21       25       6.4      4      20                                          30       25       6.1      2.5     8                                           1       50       11       30     97                                           5       50       9        12     40                                           7       50       7.5      3      10                                          10       50       6.0      1       0                                          ______________________________________                                         *Determined by titration to color neutralilty for stock solutions of          doxorubicin and mitoxantrone (1 mg/ml each). Results standardized to          initial activity.                                                             **100 ml of solution containing 4 g of calcium hypochlorite and 2.5 g of      methylcellulose.                                                         

The data of Table III show that significant activity was lost after onlyfive days of storage of the dry ingredients and that the mixture hadbecome almost completely inactive after storage for 30 days at roomtemperature or 10 days at 50° C. It is apparent, therefore, that evenwhen mixed as dry powders, the active ingredients of the mixtureinactivate each other in a manner which is dependent on time andtemperature.

EXAMPLE 5 Stability of Mixtures of Hypochlorite and Other Thickeners inAqueous Solution

In addition to methylcellulose, four common viscosity enhancing agentswere tested for suitability for increasing the viscosity of hypochloritesolutions. The results of these tests are reported in Table IV. It willbe seen that, except for methlycellulose, the other thickeners wereunsuitable for use in calcium hypochlorite solutions either because theybecame quickly deactivated (polyvinylpyrolidine and polyethylene glycol)or caused a violent reaction (agar, gelatin). Agar reacted violently andcaused the solutions to thicken to a gel.

                                      TABLE IV                                    __________________________________________________________________________    Tests of Pharmaceutical Thickeners in Calcium                                 Hypochlorite Solutions (4 g Ca(OCl).sub.2 per 100 mL)                                                  Time to Loss of                                                        Viscosity                                                                            Inactivating Ability                                 Stiffening Agent                                                                           g/100 mL                                                                           (cps)* Mitoxantrone                                                                         Doxorubicin                                   __________________________________________________________________________    Polyvinylpyrolidine                                                           40,000 Molecular Weight                                                                    14   17.8   30 min.                                                                               1 hr.                                        Polyethylene Glycol Mixture                                                   3,350 Ave. Molec. Wt.                                                                      30   18     30 min.                                                                               1 hr.                                          400 Ave. Molec. Wt.                                                                      30                                                               Agar**                                                                        (Vigorous exothermic                                                                       2.5  Too thick to                                                                         24 hr. 24 hr.                                        reaction upon mixing,                                                                           evaluate                                                    forming a gel)                                                                             5.0  Too thick to                                                                         48 hr. 48 hr.                                                          evaluate                                                                 10.0 Too thick to                                                                         --     --                                                              evaluate                                                    Gelatin      1    Impossible to evaluate due to an                                         2.5  immediate violent endothermic                                            10   reaction.                                                   __________________________________________________________________________     *Measured immediately after mixing using the falling ball method.             **The exothermic reaction precludes the use of agar in clinical settings      because of potential explosive properties.                               

EXAMPLE 6 HIV Inactivation by Dilute Calcium Hypochlorite

The ability of dilute calcium hypochlorite to inactivate HIV was testedagainst an HIV-infected CD4+ human cell line (H9/IIIB). A saturatedsolution of dilute calcium hypochlorite completely inactivated infectedhuman T-lymphocytes.

Procedure:

H9/IIIB cells (10⁶ cells/ml of complete culture medium, RPMI in 10%fetal calf serum [FCS]) were exposed to an aqueous solution containing4% by weight of calcium hypochlorite for 15 minutes; the volumetricratio of culture medium to calcium hypochlorite solution was 1:1, or a50% dilution of a 4% dilute calcium hypochlorite solution in culturemedium containing 10% serum. The cells were subsequently washed threetimes with 10% FCS RPMI and plated in fresh medium and allowed toincubate at 37° C. for three days, to allow any residual live virusand/or virus-infected cells to develop after the brief exposure to thedilute calcium hypochlorite solution. The cells were again pelleted andexposed to lytic CD4+ human cell line (MOT) as a target for any viablevirus. The target MOT cells will lyse when infected with HIV. Thesecells were then allowed to incubate an additional four days. Next, thecells were incubated with human anti-HIV antibody (prepared inaccordance the the procedures described in Lake DA, Sugano T, MatsumotoY, et al: "A Hybridoma Producing Monoclonal Antibody Specific forGlycoprotein 120 kDa of Human Immunodeficiency Virus (HIV-1), LifeSciences 45:iii-x, 1989.) for one hour, and then washed and incubatedwith fluorescent labeled goat anti-human IgG for one hour. The methodfor HIV detection by formaldehyde fixation and flow cytometry was inaccordance with the method described in Lifson JD, Sasaki DT, EnglemanEG: "Utility of Formaldehyde Fixation for Flow Cytometry andInactivation of the AIDS Associated Retrovirus", J Immunological Methods86:143-149, 1986.

Controls:

As a positive control, the above procedure was followed withHIV-infected H9/IIIB cells exposed only to culture medium rather than toculture medium and calcium hypochlorite. As a negative control, theabove procedure was followed with uninfected H9 cells exposed to anaqueous solution containing 4% by weight of calcium hypochlorite.

Results:

After a four day incubation period, the HRV positive control culturesexhibited large concentrations of swollen, HIV-infected MOT cells. Thecalcium hypochlorite solution (n=4 each) completely lysed non-infectedas well as HIV infected MOT cells, and there were no viable HIV varionspresent.

HIV infected cells exposed to dilute calcium hypochlorite were alsoviewed under a fluorescent microscope following exposure to thefluorescent anti-HIV antibody. They showed no signs of HIV infection andwere identical to the negative control.

Conclusion:

Dilute calcium hypochlorite completely inactivates humanimmunodeficiency virus when added to infected cell cultures in a 1:1volumetric ratio. The cells are completely lysed by dilute calciumhypochlorite and no HIV antibody binding is detectable using a goatantibody specific to the 120 kDa viral glycoprotein.

EXAMPLE 7 HIV Inactivation bv Dilute Calcium Hypochlorite Containing aThickener

The materials and methods of Example 6 were repeated, except that theaqueous bleach solution applied to the infected and uninfected H9 cellsincluded 2.5% by weight of the thickener methlycellulose in addition to4% by weight of calcium hypochlorite. The results were the same as thoseobtained in Example 6. The MOT cells were lysed and the residualmaterial was not infectious; however, the fluorescent entibody techniquecould not be completed because of the methlycellulose thickening effect.

EXAMPLE 8 Use of Towlette Swabs for Deactivating Anticancer DrugMaterials

Experimental Materials

Test Swabs: 17×23 cm nylon-reinforced 4 ply, #34770, each weighingapproximately 2.6 gram Terri® Wipers, manufactured by Kimberly Clark,Neenah, Wis.

Bleach: 5.25% by weight aqueous solution of household bleach, i.e.,sodium hypochlorite (NaHCl0₄), supplied by National Sanitary Supply, LosAngeles, Calif., having a pH of approximately 11.2.

Sodium thiosulfate: 5% by weight aqueous solution, supplied by TarijianLabs, Inc., Queens Village, N.Y.

Doxorubicin: 1 mg/ml aqueous solution, supplied by Adria Labs, Columbus,Ohio.

Experimental Procedure

0.1 ml of the doxorubicin solution (1 mg/ml) was applied onto a fresh,dry towelette. The doxorubicin solution caused a red colored stain toappear on the surface of the towlette. One drop of the household bleachsolution was then soueezed onto the red stained, doxorubicincontaminated towelette surface. Within 10-15 seconds the red colordisappeared, indicating that the bleach solution had chemicallyinactivated the doxorubicin. The pH of the bleach solution on thesurface of the towlette was found to be 11.2, as determined by a pHmeter.

In a glass container, 10 ml of the 5% sodium thiosulfate solution wasadded to 10 ml of the 5.25% bleach solution; a slight exothermicreaction resulted from the addition of the sodium thiosulfate to thebleach solution. Between 10-20 drops of the resulting mixture was thenadded to a second doxorubicin stain on a towelette. The red doxorubicinstain remained on the towlette, indicating that the sodium thiosulfateneutralized the bleach solution so that the bleach could no longerinactivate the doxorubicin on the towelette. A pH meter was again usedto determine the pH (1.3) of the combined sodium thiosulfate-bleachmixture on the surface of the towlette.

These results indicate that household bleach used in low volume cancause the chemical inactivation of doxorubicin droplet contamination ona paper towelettLe. Mixing together ecrual volumes of the 5% sodiumthiosulfate solution and the 5.25% household bleach solution results ina mixture incapable of decolorizing the doxorubicin contamination on thedry towelette, indicating that the sodium thiosulfate neutralized theoxidizing action of the household bleach. The pH of the combinationsodium thiosulfate/household bleach solution was 1.3.

Sodium thiosulfate increases the carcinogen inactivating capacity of thesodium hypochlorite solution due to the thiosulfate 's ability to bindchemically to various alkylating type molecules such as nitrogen mustardand cisplatin. Furthermore, the addition of sodium thiosulfate to thesodium hypochlorite solution prevents permanent etching of stainless andceramic surfaces caused by the alkaline sodium hypochlorite solution.

Various combinations and pHs of acid alcohol (a mixture of absoluteethanol and 0.1 N hydrochloric acid), mixed with sodium hypochloriteand/or sodium thiosulfate, were evaluated as listed below.

In a series of experiments, between 1-10 ml acid alcohol, having pHs inthe range of between 1-3.75, were added to between 5-10 ml householdbleach. The acid alcohol was unable to inactivate the household bleachuntil the ratio of acid alcohol to bleach (having a pH of 1.0) was 2:1;at that high ratio of acid alcohol to bleach, a moderate exothermicreaction as well as noxious fumes occurred.

In another series of experiments, between 2.5-10 ml sodium thiosulfate,5-10% were added to between 0-9 ml ethanol, together with 10 mlhousehold bleach. The mixture of the three chemicals resulted inmoderately severe exothermic reactions and inactivation of bleach'soxidizing activity only when equal ratios of 5% sodium thiosulfate wereadded to the bleach (in the presence of ethanol), and the resulting pHwas below 2.

The foregoing description has been given for clearness of understandingonly, and no unnecessary limitations should be understood therefrom, asmodifications will be obvious to those skilled in the art.

EXAMPLE 9 Use of Towlette Swabs for Deactivating Work SurfacesContaminated by An Anticancer Drug Material

The Experimental Materials used are as described above in Example 8. Afirst fresh, dry test swab or towlette is impregnated with a 5.25% byweight aqueous solution of sodium hypochlorite by applying 5 (five) ml.of the sodium hypochlorite solution to the test swab. A second fresh,dry test swab or towlette is impregnated with a 5% by weight aqueoussolution of sodium thiosulfate by applying 5 (five) ml. of the sodiumthiosulfate solution to the test swab.

Doxorubicin is placed on a stainless steel work surface, which shouldcause a red stain to appear. The sodium hypochlorite impregnatedtowlette is then wiped or swabbed, over the doxorubicin spill, whichshould cause the red stain to disappear, indicating chemicalinactivation of the doxorubicin by the sodium hypochlorite.

The sodium thiosulfate impregnated towlette is then wiped, or swabbed,over the work surface containing the chemically inactivated doxorubicinspill. The sodium thiosulfate should neutralize any residual sodiumhypochlorite left on the work surface, thereby preventing the stainlesssteel surface from being etched by the residual alkaline sodiumhypochlorite solution.

EXAMPLE 10 Use of Towlette Swabs for Deactivating Work SurfacesContaminated bv HIV Infected Cells

Experimental Materials

Test Swabs: 17×23 cm nylon-reinforced 4 ply, #34770, each weighingapproximately 2.6 gram Terri® Wipers, manufactured by Kimberly Clark,Neenah, Wis.

Bleach: 4% by weight aqueous solution of calcium hypochlorite.

Sodium thiosulfate: 5% by weight aqueous solution, supplied by TarijianLabs, Inc., Queens Village, N.Y.

HIV infected Cells: H9/IIIB cells (10⁶ cells/ml of complete culturemedium, RPI in 10% fetal calf serum [FCS]), as described above inExample 6.

A first fresh, dry test swab or towlette is impregnated with a 4.0% byweight aqueous solution of calcium hypochlorite by applying 5 (five) ml.of the calcium hypochlorite solution to the test swab. A second fresh,dry test swab or towlette is impregnated with a 5% by weight aqueoussolution of sodium thiosulfate by applying 5 (five) ml. of the sodiumthiosulfate solution to the test swab.

A small amount of the HIV infected cells are placed on a stainless steelwork surface. The sodium hypochlorite impregnated towlette is then wipedor swabbed, over the HIV infected cells, which should inactivate thevirus.

The sodium thiosulfate impregnated towlette is then wiped, or swabbed,over the work surface containing the inactivated virus. The sodiumthiosulfate should neutralize any residual calcium hypochlorite left onthe work surface, thereby preventing the stainless steel surface frombeing etched by the residual alkaline calcium hypochlorite solution.

The foregoing description has been given for clearness of understandingonly, and no unnecessary limitations should be understood therefrom, asmodifications will be obvious to those skilled in the art.

What is claimed is:
 1. A method for inactivating HIV infected blood, themethod comprising the steps of:(a) swabbing a surface containing the HIVinfected blood with a first absorbent, fibrous towlette impregnated withan aqueous solution of a deactivator selected from the group consistingof calcium hypochlorite, sodium hypochlorite and mixtures thereof, toinactivate HIV present in such HIV infected blood; and (b) thereafterswabbing the surface with a second absorbent, fibrous towletteimpregnated with an aqueous solution of sodium thiosulfate to neutralizeany residual deactivator present on the surface so as to prevent etchingof the surface by the residual deactivator.
 2. The method of claim 1wherein the first and second towlettes are made from materials selectedfrom the group consisting of cotton, synthetic blends, and mixturesthereof, the deactivator is sodium hypochlorite, and wherein the ratioof deactivator volume to first towlette weight is in the range of 1.8 to15 ml/gm, and the ratio of sodium thiosulfate volume to second towletteweight is in the range of 1.8 to 15 ml/gm.
 3. A method for inactivatingan anticancer drug selected from the group of anticancer drugsconsisting of L-asparaginase, carmustine, doxorubicin, daunorubicin,epirubicin, mitoxantrone, mitomycin-C, melphalan, cyclophosphamide,ifosphamide, thio-TEPA, decarbazine, cisplatin, carboplatin,vincristine, vinblastine, vindesine, actinomycin D, bleomycin,etoposide, teniposide, α-interferon, interleukin 2, tumor necrosisfactor, G-CSF, and GM-CSF, the method comprising the steps of:(a)swabbing a surface containing the anticancer drug with a firstabsorbent, fibrous towlette impregnated with an aqueous solution of adeactivator selected from the group consisting of calcium hypochlorite,sodium hypochlorite and mixtures thereof, to inactivate the anticancerdrug; and (b) thereafter swabbing the surface with a second absorbent,fibrous towlette impregnated with an aqueous solution of sodiumthiosulfate to neutralize any residual deactivator present on thesurface so as to prevent etching of the surface by the residualdeactivator.
 4. The method of claim 3 wherein the first and secondtowlettes are made from materials selected from the group consisting ofcotton, synthetic blends, and mixtures thereof, the deactivator issodium hypochlorite, and wherein the ratio of deactivator volume tofirst towlette weight is in the range of 1.8 to 15 ml/gm, and the ratioof sodium thiosulfate volume to second towlette weight is in the rangeof 1.8 to 15 ml/gm.